Abstract
Brain vessels are the most important structures in the brain to deliver energy and substrates to neurons. Brain vessels are composed of a complex interaction between endothelial cells, pericytes, and astrocytes, controlling the entry of substrates into the brain. Damage of brain vessels and vascular impairment are general pathologies observed in different neurodegenerative disorders including e.g., Alzheimer's disease. In order to study remodeling of brain vessels, simple 3-dimensional in vitro systems need to be developed. Organotypic brain slices of mice provide a potent tool to explore angiogenic effects of brain vessels in a complex 3-dimensional structure. Here we show that organotypic brain slices can be cultured from 110 μm thick sections of postnatal and adult mice brains. The vessels are immunohistochemically stained for laminin and collagen IV. Co-stainings are an appropriate method to visualize interaction of brain endothelial cells with pericytes and astrocytes in these vessels. Different exogenous stimuli such as fibroblast growth factor-2 or vascular endothelial growth factor induce angiogenesis or re-growth, respectively. Hyperthermia or acidosis reduces the vessel density in organotypic slices. In conclusion, organotypic brain slices exhibit a strong vascular network which can be used to study remodeling and angiogenesis of brain vessels in a 3-dimensional in vitro system.
Highlights
The organotypic brain slice model resembles partly the in vivo condition of a high density cell system
In the present work we summarize published experiments and present examples of own data on exogenous modulation of the vascular network in organotypic brain slices
We show that brain slices exhibit a dense vascular network and are positive for laminin and collagen IV
Summary
The organotypic brain slice model resembles partly the in vivo condition of a high density cell system. We were one of the first to demonstrate that organotypic brain slices contain a strong network of laminin+ brain capillaries (Moser et al, 2003, 2004). We have recently shown that brain vessels in organotypic brain slices can re-grow between specific areas, when exogenously stimulated (Ullrich and Humpel, 2009). Brain vessels are well studied in organotypic cultures and the neurovascular unit (NVU) is intensively explored in this complex 3-dimensional network (Morin-Brureau et al, 2013; Chip et al, 2014). The interaction of vascular cells with astrocytes and with pericytes within 3-dimensional organotypic slices came into intense investigation (Mishra et al, 2014). The interaction of vascular cells with astrocytes and with pericytes within 3-dimensional organotypic slices came into intense investigation (Mishra et al, 2014). Zehendner et al (2013) provided a detailed characterization of such a novel organotypic in vitro model of the NVU in the developing cortex
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